scholarly journals DNA Authentication of St John’s Wort (Hypericum perforatum L.) Commercial Products Targeting the ITS Region

Genes ◽  
2019 ◽  
Vol 10 (4) ◽  
pp. 286
Author(s):  
Howard ◽  
Hill ◽  
Kreuzer ◽  
Mali ◽  
Masiero ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Hypericum Perforatum ◽  
Its Region ◽  
Qpcr Assay ◽  
Plant Materials ◽  
Commercial Products ◽  
Synthetic Dna ◽  
Hypericum Perforatum L ◽  
Species Specific ◽  
Its1 And Its2

There is considerable potential for the use of DNA barcoding methods to authenticate raw medicinal plant materials, but their application to testing commercial products has been controversial. A simple PCR test targeting species-specific sequences within the nuclear ribosomal internal transcribed spacer (ITS) region was adapted to screen commercial products for the presence of Hypericum perforatum L. material. DNA differing widely in amount and extent of fragmentation was detected in a number of product types. Two assays were designed to further analyse this DNA using a curated database of selected Hypericum ITS sequences: A qPCR assay based on a species-specific primer pair spanning the ITS1 and ITS2 regions, using synthetic DNA reference standards for DNA quantitation and a Next Generation Sequencing (NGS) assay separately targeting the ITS1 and ITS2 regions. The ability of the assays to detect H. perforatum DNA sequences in processed medicines was investigated. Out of twenty different matrices tested, both assays detected H. perforatum DNA in five samples with more than 103 ITS copies µL−1 DNA extract, whilst the qPCR assay was also able to detect lower levels of DNA in two further samples. The NGS assay confirmed that H. perforatum was the major species in all five positive samples, though trace contaminants were also detected.

scholarly journals A Forensic Detection Method for Hallucinogenic Mushrooms via High-Resolution Melting (HRM) Analysis

Genes ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 199
Author(s):  
Xiaochun Zhang ◽  
Huan Yu ◽  
Qi Yang ◽  
Ziwei Wang ◽  
Ruocheng Xia ◽  
...  
Keyword(s):  
High Resolution ◽  
Dna Barcoding ◽  
High Resolution Melting ◽  
Its Region ◽  
Its Sequencing ◽  
Melting Temperatures ◽  
Hrm Analysis ◽  
National Drug ◽  
Species Specific ◽  
Its1 And Its2

In recent years, trafficking and abuse of hallucinogenic mushrooms have become a serious social problem. It is therefore imperative to identify hallucinogenic mushrooms of the genus Psilocybe for national drug control legislation. An internal transcribed spacer (ITS) is a DNA barcoding tool utilized for species identification. Many methods have been used to discriminate the ITS region, but they are often limited by having a low resolution. In this study, we sought to analyze the ITS and its fragments, ITS1 and ITS2, by using high-resolution melting (HRM) analysis, which is a rapid and sensitive method for evaluating sequence variation within PCR amplicons. The ITS HRM assay was tested for specificity, reproducibility, sensitivity, and the capacity to analyze mixture samples. It was shown that the melting temperatures of the ITS, ITS1, and ITS2 of Psilocybe cubensis were 83.72 ± 0.01, 80.98 ± 0.06, and 83.46 ± 0.08 °C, and for other species, we also obtained species-specific results. Finally, we performed ITS sequencing to validate the presumptive taxonomic identity of our samples, and the sequencing output significantly supported our HRM data. Taken together, these results indicate that the HRM method can quickly distinguish the DNA barcoding of Psilocybe cubensis and other fungi, which can be utilized for drug trafficking cases and forensic science.

scholarly journals Sequence Analysis of the Internal Transcribed Spacer (ITS) Region of the Nuclear Ribosomal DNA (nrDNA) and Chloroplast trnL-F Region (cpDNA) of Some Lactuca L. (Asteraceae) Species in Turkey

2016 ◽  
Vol 8 (4) ◽  
pp. 444-450 ◽  
Author(s):  
Emre SEVİNDİK ◽  
Veysel UZUN ◽  
Fatih COŞKUN
Keyword(s):  
Sequence Analysis ◽  
Dna Sequences ◽  
Phylogenetic Trees ◽  
Its Region ◽  
Nucleotide Composition ◽  
Nuclear Ribosomal Dna ◽  
Sequencing Data ◽  
Plant Materials ◽  
F Region ◽  
Lactuca Species

In the current study, sequence analysis of some Turkish Lactuca L. species using nrITS DNA and trnL-F cpDNA sequences were performed to elucidate phylogenetic relationships among the taxa under study. Hieracium umbellatum was used as an outgroup. Different plant materials of Lactuca were collected from different parts of Turkey during excursions of summer 2013. Plant materials were either kept in silica gel or kept fresh for immediate DNA isolation. Both phenol chloroform-isoamyl alcohol method and commercial kits were used to extract genomic DNA for PCR reactions. ITS4 and ITS5A primers were utilized for ITS region, while trnLe and trnLf primers were used to amplify the trnL-F region. Obtained DNA sequences were edited both manually and by using BioEdit 7.0.4.1. Sequencing data were aligned via ClustalW program and analyzed using PAUP 4.01b10 software. nrITS sequences varied from 639 nucleotides to 735 nucleotides. Average nucleotide composition for nrITS was 22.1% (T), 27.9% (C), 23.2% (A) and 26.8% (G). It was also found that divergence values differed between 0.0000 and 0.10290. The trnL-F sequences varied from 296 nucleotides to 385 nucleotides. Average nucleotide composition of trnL-F sequences was 34.1% (T), 18.4% (C), 31.6% (A) and 16.0% (G). It was also found that divergence values differed between 0.0000 and 0.09674. Neighbour Joining (NJ) trees were constructed in order to identify the relationships among Lactuca species. Phylogenetic trees based on ITS region were found to be more useful than phylogenetic trees based on trnL-F region. After analysis of the results obtained, the data suggest that Lactuca contains 2 clades, with clade 1 having 2 subclades. These results support the prior phylogenetic studies on Lactuca and hence provide an up to date review of Turkish Lactuca species.

Assessment of cadmium intake from commercial products based on herb St. Johns wort ( Hypericum perforatum L.)

2015 ◽  
Vol 238 (2) ◽  
pp. S130 ◽  
Author(s):  
D. Djukic-Cosic ◽  
A. Buha ◽  
Z. Bulat ◽  
A. Ugarkovic ◽  
V. Berta ◽  
...  
Keyword(s):  
Hypericum Perforatum ◽  
Commercial Products ◽  
Cadmium Intake ◽  
Hypericum Perforatum L

Molecular and morphological characterisation of Scutellonema bradys from yam in Costa Rica and development of specific primers for its detection

Nematology ◽  
2014 ◽  
Vol 16 (2) ◽  
pp. 137-147 ◽  
Author(s):  
Danny A. Humphreys-Pereira ◽  
Valerie Moroz Williamson ◽  
Sooung Lee ◽  
Daniel L. Coyne ◽  
Luis Salazar ◽  
...  
Keyword(s):  
Costa Rica ◽  
Dna Sequences ◽  
Its Region ◽  
Geographic Region ◽  
Specific Primers ◽  
Dry Rot ◽  
Male Characters ◽  
Yellow Yam ◽  
Rot Disease ◽  
Species Specific

The yam nematode, Scutellonema bradys, which can cause dry rot disease of yam (Dioscorea spp.), was recorded for the first time from Costa Rica in four species of yam occurring in the Atlantic and north regions. Morphometric measurements from two populations from each region using ten female and 11 male characters corresponded with previous descriptions of this species. Canonical discriminant analysis of the female morphometric data separated the populations by region, whereas no separation by region was evident using the male data. Analysis of DNA sequences from the ITS region indicated that populations from Costa Rica were monophyletic with S. bradys from West Africa and clearly distinct from other Scutellonema species. No genetic separation by geographic region or Dioscorea species host was observed between Costa Rica populations. Species-specific primers were developed from the ITS region and supported the identity of 17 populations from 15 locations in Costa Rica as S. bradys: 14 populations from D. alata (greater or water yam) and one each from D. trifida (white yampee), D. cayenensis (yellow yam) and D. rotundata (white yam). Yam production in Costa Rica began in the Atlantic region, where the yam nematode was likely introduced from the Caribbean, progressively spreading to other locations through the use of infected vegetative planting material.

scholarly journals Applied Barcoding: The Practicalities of DNA Testing for Herbals

Plants ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 1150 ◽  
Author(s):  
Caroline Howard ◽  
Claire Lockie-Williams ◽  
Adrian Slater
Keyword(s):  
Dna Sequences ◽  
Dna Barcode ◽  
Plant Materials ◽  
Quality Testing ◽  
Hypericum Perforatum L ◽  
Public Repositories ◽  
Testing Protocols ◽  
Significant Attention ◽  
Hypericum Species

DNA barcoding is a widely accepted technique for the identification of plant materials, and its application to the authentication of commercial medicinal plants has attracted significant attention. The incorporation of DNA-based technologies into the quality testing protocols of international pharmacopoeias represents a step-change in status, requiring the establishment of standardized, reliable and reproducible methods. The process by which this can be achieved for any herbal medicine is described, using Hypericum perforatum L. (St John’s Wort) and potential adulterant Hypericum species as a case study. A range of practical issues are considered including quality control of DNA sequences from public repositories and the construction of individual curated databases, choice of DNA barcode region(s) and the identification of informative polymorphic nucleotide sequences. A decision tree informs the structure of the manuscript and provides a template to guide the development of future DNA barcode tests for herbals.

Determination of Hypericin in St. John's Wort (Hypericum perforatum L.) extracts using HPLC-ED

Planta Medica ◽  
2011 ◽  
Vol 77 (12) ◽  
Author(s):  
E Sofic ◽  
A Copra Janicijevic ◽  
M Maksimovic ◽  
I Tahirovic ◽  
L Klepo ◽  
...  
Keyword(s):  
Hypericum Perforatum ◽  
St John’S Wort ◽  
St John's Wort ◽  
Hypericum Perforatum L
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